microbial structure and function mi720 fall 2007 dna replication andrew j. pierce department of...
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![Page 1: Microbial Structure and Function MI720 Fall 2007 DNA Replication Andrew J. Pierce Department of Microbiology, Immunology and Molecular Genetics University](https://reader035.vdocuments.site/reader035/viewer/2022062314/56649d9f5503460f94a8a89c/html5/thumbnails/1.jpg)
Microbial Structure and FunctionMI720
Fall 2007
DNA Replication
Andrew J. Pierce
Department of Microbiology, Immunology and Molecular Genetics
University of Kentucky
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FEMS Microbiol Rev.2002 Nov;26(4):355-74.The bacterial replication initiator DnaA. DnaA and oriC, the bacterial mode to initiate DNA replication.Messer W.
E. coli origin structure
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DnaA boxes are shown as red arrowsAT-rich regions are shown in yellow
Replication origins of different bacteria
FEMS Microbiol Rev.2002 Nov;26(4):355-74.The bacterial replication initiator DnaA. DnaA and oriC, the bacterial mode to initiate DNA replication.Messer W.
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FEMS Microbiol Rev.2002 Nov;26(4):355-74.The bacterial replication initiator DnaA. DnaA and oriC, the bacterial mode to initiate DNA replication.Messer W.
5'-TTA/TTNCACA-3'
A-box: KD ~25 nM
initial unwinding: DnaA: 28 bpDnaA+SSB: 44-52 bp
Initial origin melting
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Origin proteins
dnaA melt duplex
dnaB main replicative helicasecommitment step for replication fork formation
dnaC load dnaB
dnaG primase
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FEMS Microbiol Rev.2002 Nov;26(4):355-74.The bacterial replication initiator DnaA. DnaA and oriC, the bacterial mode to initiate DNA replication.Messer W.
Initiating DNA replication
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Annu Rev Biochem. 1995;64:171-200DNA polymerase III holoenzyme: structure and function of a chromosomal replicating machineKelman Z, O'Donnell M.
2 DNA polymerase molecules and 9 other accessory factors
E. coli DNA polymerase III holoenzyme
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Principles of Biochemistry, 3rd EditionHorton HR, Moran LA, Ochs RS, Rawn DJ, Scrimgeour KGPrentice Hall 2002
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replicationfork
replicationfork
Principles of Biochemistry, 3rd EditionHorton HR, Moran LA, Ochs RS, Rawn DJ, Scrimgeour KGPrentice Hall 2002
Replicating E. coli genome
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Other E. coli DNA polymerases
DNA pol I remove RNA primers and join Okazaki fragments
DNA pol II aka: dinA (damage inducible), polBproofread terminal mismatchesextend terminal mismatches
DNA pol IV aka: dinB (damage inducible)extend terminal mismatches without proofreading
DNA pol V aka: umuD'2Ctranslesion synthesis
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When Pol III goes wrong…mismatches
Mol Microbiol.2005 Oct;58(1):61-70DNA polymerase II as a fidelity factor in chromosomal DNA synthesis in Escherichia coliBanach-Orlowska M, Fijalkowska IJ, Schaaper RM, Jonczyk P
pol IV also used for template slippage
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Trends Biochem Sci.2000 Apr;25(4):189-95Coping with replication 'train wrecks' in Escherichia coli using PolV, Pol II and RecA proteinsGoodman MF.
Bigger problems for DNA replication
abasicsite
UV 6-4 photoproduct UV cyclobutane dimer
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SOS response
TTRecA
RecA *
RecA* LexA
Genetics, Vol. 158, 41-64, May 2001Comparative gene expression profiles following UV exposure in wild-type and SOS-deficient Escherichia coliCourcells J, Khodursky A, Peter B, Borwn PO, Hanawalt PC
uvrA nucleotideuvrB excisionuvrD repair
dinA = polB = pol IIdinB = pol IV
sulA block cell division
lexArecA
ruvA resistanceruvB to UV
umuC UVumuD'umuD mutagenesis
RecA*
1 2
3
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Proc Natl Acad Sci USA2001 Jul 17;98(15):8350-4Roles of DNA polymerases V and II in SOS-induced error-prone and error-free repair in Escherichia coliPham P, Rangarajan S, Woodgate R, Goodman MF
Pol II helping out
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Proc Natl Acad Sci USA2001 Jul 17;98(15):8350-4Roles of DNA polymerases V and II in SOS-induced error-prone and error-free repair in Escherichia coliPham P, Rangarajan S, Woodgate R, Goodman MF
Translesion synthesis
polV: umuD'2C
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E. coli genome
Microbiol Mol Biol Rev. 2005 Sep;69(3):501-26Replication termination in Escherichia coli: structure and antihelicase activity of the Tus-Ter complexNeylon C, Kralicek AV, Hill TM, Dixon NE
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Mol Microbiol. 1999 Mar;31(6):1611-8Termination of DNA replication of bacterial and plasmid chromosomesBussiere DE, Bastia D
Replication termination
TerF TerB TerC TerA TerD TerE
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Tus - Ter
Mol Microbiol. 1999 Mar;31(6):1611-8Termination of DNA replication of bacterial and plasmid chromosomesBussiere DE, Bastia D
5'-TTAGTTACAACATNCA-3'
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Tus - Ter and DNA polymerase III
Microbiol Mol Biol Rev. 2005 Sep;69(3):501-26Replication termination in Escherichia coli: structure and antihelicase activity of the Tus-Ter complexNeylon C, Kralicek AV, Hill TM, Dixon NE
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Replication fork disassembly
Microbiol Mol Biol Rev. 2005 Sep;69(3):501-26Replication termination in Escherichia coli: structure and antihelicase activity of the Tus-Ter complexNeylon C, Kralicek AV, Hill TM, Dixon NE